# Increased Nucleosomes and Neutrophil Activation Link to Disease Progression in Patients with Scrub Typhus but Not Murine Typhus in Laos

## TL;DR

The study shows that high levels of nucleosomes and neutrophil activation are linked to severe disease in scrub typhus patients in Laos, but not in those with murine typhus.

## Contribution

The study identifies nucleosomes and neutrophil activation as independent risk factors for severe scrub typhus, not observed in murine typhus.

## Key findings

- Elevated nucleosome levels correlate with severe symptoms and liver dysfunction in scrub typhus.
- Neutrophil activation markers like ELA complexes are linked to fibrinolysis and severe disease progression in scrub typhus.
- Nucleosomes and ELA complexes are strong predictors of severe scrub typhus, with 4- to 5-fold increased risk.

## Abstract

Cell-mediated immunity is essential in protection against rickettsial illnesses, but the role of neutrophils in these intracellular vasculotropic infections remains unclear. This study analyzed the plasma levels of nucleosomes, FSAP-activation (nucleosome-releasing factor), and neutrophil activation, as evidenced by neutrophil-elastase (ELA) complexes, in sympatric Lao patients with scrub typhus and murine typhus. In acute scrub typhus elevated nucleosome levels correlated with lower GCS scores, raised respiratory rate, jaundice and impaired liver function, whereas neutrophil activation correlated with fibrinolysis and high IL-8 plasma levels, a recently identified predictor of severe disease and mortality. Nucleosome and ELA complex levels were associated with a 4.8-fold and 4-fold increased risk of developing severe scrub typhus, beyond cut off values of 1,040 U/ml for nucleosomes and 275 U/ml for ELA complexes respectively. In murine typhus, nucleosome levels associated with pro-inflammatory cytokines and the duration of illness, while ELA complexes correlated strongly with inflammation markers, jaundice and increased respiratory rates. This study found strong correlations between circulating nucleosomes and neutrophil activation in patients with scrub typhus, but not murine typhus, providing indirect evidence that nucleosomes could originate from neutrophil extracellular trap (NET) degradation. High circulating plasma nucleosomes and ELA complexes represent independent risk factors for developing severe complications in scrub typhus. As nucleosomes and histones exposed on NETs are highly cytotoxic to endothelial cells and are strongly pro-coagulant, neutrophil-derived nucleosomes could contribute to vascular damage, the pro-coagulant state and exacerbation of disease in scrub typhus, thus indicating a detrimental role of neutrophil activation. The data suggest that increased neutrophil activation relates to disease progression and severe complications, and increased plasma levels of nucleosomes and ELA complexes represent independent risk factors for developing severe scrub typhus.

Tropical rickettsial illnesses, especially scrub typhus and murine typhus, are increasingly recognized as a leading cause of treatable undifferentiated febrile illness in Asia, but remain severely neglected and under appreciated diseases in many areas. In this study we investigated the relationship of markers of neutrophil activation and cell death with disease severity in patients with acute scrub typhus and murine typhus in Laos. These easily measurable circulating markers were associated with a 4 to 5-fold increased risk of developing severe clinical disease manifestations in scrub typhus and represent independent predictors of severe disease, and possibly death. We also found strong correlations between circulating markers of cell death and neutrophil activation in patients with scrub typhus, but not murine typhus, providing indirect evidence that neutrophil extracellular traps could contribute to the vascular damage and pro-coagulant state leading to exacerbation of disease in scrub typhus, thus indicating a detrimental role of neutrophil activation. The data suggest that increased neutrophil activation relates to disease progression and severe complications, and increased plasma levels of nucleosomes and ELA complexes represent independent risk factors for developing severe scrub typhus.